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. 2019 Jan 18;9:208. doi: 10.1038/s41598-018-36312-4

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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sumo-1/2/3 and SUMO-1/2/3 levels increased as LMP1 levels increased in EBV-positive cell lines. (a) RNA was extracted from paired naïve lymphocytes, mitogen blasts, and EBV-transformed LCLs. cDNAs were generated, and real-time PCR was performed and relative sumo-1, and sumo-2/3 levels (relative to gapdh) determined. Results are shown as the mean ± the standard deviation for samples performed in triplicate. Independent experiments were performed in triplicate. (b–e) RNA and protein were harvested from ten different EBV-positive cell lines. (c) cDNAs were generated, and real-time PCR was performed to quantitate relative LMP1, sumo-1, and sumo-2/3 levels (relative to gapdh). Results are shown as the mean for samples performed in triplicate from three independent experiments. Regression analysis of LMP1 and sumo-1 or sumo-2/3 levels was performed. (d–f) Slot immunoblots were performed to detect LMP1, SUMO-1, and SUMO-2/3 levels in (d) three EBV-transformed LCLs, (e) two KR4-HeLa fusion cell lines, and (f) three EBV-infected human breast cancer cell lines. GAPDH was used as a loading control. Representative blots for experiments performed in triplicate are shown.